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Digestible substrates for cell culture

A cell culture and substrate technology, applied in cell culture supports/coatings, tissue culture, animal cells, etc., to solve problems such as vaccines that cannot be used for anti-cancer treatment

Inactive Publication Date: 2018-03-27
CORNING INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] As another example of the detrimental effects of proteases, treatment of cells with proteases such as trypsin is also known to remove antigens from cancer cells, thus potentially rendering them unusable for the development of vaccines for anticancer treatments

Method used

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  • Digestible substrates for cell culture
  • Digestible substrates for cell culture
  • Digestible substrates for cell culture

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0083] Example 1. 1% PGA microbeads cross-linked with 3% calcium.

[0084] Microgalacturonic acid (PGA) solutions were prepared from 1% by weight polygalacturonic acid (PGA) solutions by dissolving polygalacturonic acid sodium salt [Sigma Cat. beads. Filter the solution under vacuum using a 20 micron polypropylene filter to remove particles in suspension.

[0085] A gel bath was prepared using 400 ml of a 3% (w / v) calcium chloride solution in water / ethanol (75 / 25 v / v) in a separate beaker, which was stirred using a magnetic stirrer.

[0086] Droplets were prepared by adding 25 ml of the PGA solution to the gel bath using a syringe fitted with a 30 gauge needle. A syringe pressure of about 2 bar is applied.

[0087] The beads were hardened in a calcium chloride bath for 120 minutes, followed by four washes with water. The calcium content in the beads was determined as described in Example 9. After four washes, the calcium concentration was about 0.5-0.6 g / l wetted beads....

Embodiment 2

[0090] Example 2. 1% PGA microcarriers cross-linked with 12% calcium.

[0091] The procedure of Example 1 was repeated except that 12% (w / v) calcium chloride in water / ethanol (75 / 25 v / v) solution was used instead of 3% (w / v) calcium chloride in water / ethanol (75 / 25 v / v) solution. After four washes, the calcium concentration was about 0.5-0.6 g / l wetted beads. The beads were highly transparent and free from any observable surface defects.

[0092] When contacted with 5mM EDTA / 50U pectinase at 25°C, the beads completely dissolved within 5 minutes.

Embodiment 3

[0093] Example 3. 1.5% PGA microcarriers cross-linked with 3% calcium.

[0094] The procedure of Example 1 was repeated, except that a 1.5% by weight solution of polygalacturonic acid and a pressure of 4 bar were used.

[0095] After four washes, the calcium concentration was about 0.7-0.8 g / l wetted beads. The beads were highly transparent and free from any observable surface defects.

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Abstract

A cell culture article is provided. The cell culture substrate includes a polygalacturonic acid compound selected from at least one of: pectic acid or salts thereof, and partially esterified pectic acid having a degree of esterification from 1 to 40 mol% or salts thereof. The polygalacturonic acid compound is crosslinked with a divalent cation and the divalent cation concentration ranges from 0.5to 2 g / l of the substrate.

Description

[0001] Cross References to Related Applications [0002] This application claims the benefit of priority under 35 U.S.C. §119 to U.S. Provisional Application Serial No. 62 / 233,044, filed September 25, 2015, and U.S. Provisional Application Serial No. 62 / 172,299, filed June 8, 2015 , this application is based on the contents of which is incorporated herein by reference in its entirety. This application is based upon the contents of commonly assigned U.S. Patent Nos. 8,4044,85 and 8,426,176 and co-pending and commonly assigned International Applications Nos. WO2014 / 209865 and WO2014 / 0120616, It is hereby incorporated by reference in its entirety. [0003] background technical field [0004] The present disclosure generally relates to methods of making digestible substrates, and more specifically, to transparent digestible microcarriers that can be used, for example, to isolate proteins, cells, and viruses, and also for diagnostic applications and cell culture. Background tec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/00C12N5/0775C08L5/00
CPCC08L5/00C12N5/0075C12N5/0663C12N2531/00C12N5/0686C12N5/0688C12N2533/18C12N2533/70
Inventor S·J·卡拉西D·亨利C·威尔莱克周悦
Owner CORNING INC
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